Modem medical science is constantly searching for new and more powerful agents to prevent, treat or retard bacterial and viral infections and cure the diseases they cause. Bacterial and viral infections of humans and domestic animals cost billions of dollars annually. Vast sums of money are spent each year by pharmaceutical companies to identify, characterize, and produce new antibiotics and antivirals to combat the emerging drug resistant strains which have become a serious problem. Reliable prophylactic treatments for disease prevention are also of major interest. Yet, despite the costs and efforts to identify treatments for viral infections, such as hepatitis and AIDS, effective therapies remain elusive.
Hepatitis is a disease of the human liver. It is manifested with inflammation of the liver and is usually caused by viral infections and sometimes from toxic agents. Hepatitis may progress to liver cirrhosis, liver cancer, and eventually death. Several viruses such as hepatitis A, B, C, D, E and G are known to cause various types of viral hepatitis. Among them, HBV and HCV are the most serious. HBV is a DNA virus with a virion size of 42 nm. HCV is a RNA virus with a virion size of 30-60 nm. See D. S. Chen, J. Formos. Med. Assoc., 95(1), 6-12 (1996).
Hepatitis B is a major health problem worldwide, especially in Asia and Africa. Approximately 300 million people are chronically infected with HBV worldwide. More than one million carriers of HBV are found in the United States. HBV infection is currently the main cause of liver cirrhosis and cancer. HBV carriers are not only long-term reservoirs of the virus but also may develop chronic liver disease and have a greatly increased risk of developing liver cirrhosis and cancer. The progression from chronic hepatitis B to cirrhosis is frequently insidious and occurs without a noticeable change in symptoms. Once the symptoms of cirrhosis or cancer are manifested, therapies are of little value.
Prevention of HBV infection is possible through vaccination which is safe and effective. However, vaccination is not effective in treating those already infected, i.e., carriers and patients. Many drugs have been used in treating chronic hepatitis B and none have been proven to be effective, except interferon. Treatment with interferon has limited success and has frequently associated with adverse side effects such as fatigue, fever, chills, headache, myalgias, arthralgias, mild alopecia, psychiatric effects and associated disorders, autoimmune phenomena and associated disorders and thyroid dysfunction. Treatment with interferon for sixteen (16) weeks has been shown to be effective with a sustained loss of viral replication in approximately 40% of hepatitis B patients. The great majority of responders had normal serum aminotransferase levels and relapse rates appeared to be low. See R. P. Perrillo, Digestive Diseases and Sciences, 38(4), 577-593 (1993). However, a higher long-term relapse rate (24%) was reported in Chinese patients with chronic hepatitis B who underwent interferon therapy. See A. S. F. Lok, H. T. Chung, V. W. S. Liu, & O. C. K. Ma, Gastroenterology, 105(6), 1833-1838 (1993).
Moreover, serum hepatitis B surface antigen (HBsAg) disappeared in 10 to 15% of patients treated with interferon. The loss of HBsAg coincided with the disappearance of HBV. Improvement in liver histology was sustained years later in HBsAg-negative patients. The lack of disease progression could thus conceivably result in the prevention of liver cancer when treatment is provided in the pre-cirrhotic stage of infection. See R. P. Perrillo, Digestive Diseases and Sciences, 38(4), 577-593 (1993).
Hepatitis C has been previously described as a non-A non-B hepatitis, which is caused by HCV. There are approximately 100 million HCV carriers worldwide. An estimated 3.5 million people have chronic hepatitis C in the United States. HCV infection will lead to liver cirrhosis and cancer with less clinical manifestation. Most hepatitis C patients do not have particular symptoms and can thus be easily overlooked until it is too late for therapy. This poses a potentially more serious problem than hepatitis B. HCV carriers also become long-term reservoirs of the virus and eventually develop chronic liver disease and have a greatly increased risk of developing liver cirrhosis and cancer. See D. S. Chen, Science, 262, 369-370 (1993).
No effective immunization is currently available, and hepatitis C can only be controlled by preventive measures such as improvement in hygiene and sanitary conditions and interrupting the route of transmission. At present, the only acceptable treatment for chronic hepatitis C is interferon which requires at least six (6) months of treatment. Initial treatment has a response rate of about 50%. However, half of those responding relapse after cessation of interferon treatment. Therefore, only about 25% of patients have a sustained response. See D. S. Chen, J. Formos. Med Assoc., 95(1), 6-12 (1996) and N. Terrault & T. Wright, New Engl. J. Med., 332(22), 1509-1511 (1995). Because the interferon therapy has limited efficacy and frequent adverse effects, a more effective regimen is needed.
AIDS is a deadly disease caused by HIV. It has been plaguing the world since the first description of the disease in 1981 and the discovery of its causative agent, HIV, in 1983. About 13 million people were infected with HIV worldwide in 1993 and the number has increased to about 21 million in 1996. See B. Jasny, Science, 260(5112), 1219 (1993) and P. Piot, Science, 272(5270), 1855 (1996).
Several drugs have been approved for treatment of this devastating disease, such as azidovudine (AZT), didanosine (dideoxyinosine, ddI), d4T, zalcitabine (dideoxycytosine, ddC), nevirapine, lamivudine (epivir, 3TC), saquinavir (Invirase), ritonavir (Norvir), indinavir (Crixivan), and delavirdine (Rescriptor). See M. I. Johnston & D. F. Hoth, Science, 260(5112), 1286-1293 (1993) and D. D. Richman, Science, 272(5270), 1886-1888 (1996
All drugs currently approved for AIDS treatment utilize inhibition of viral proliferation and are viral reverse transcriptase inhibitors or viral protease inhibitors. More protease inhibitors, such as nelfinavir and improved saquinavir, are in development. An AIDS vaccine (Salk's vaccine) has been tested and several proteins which are chemokines from CD8 have been discovered to act as HIV suppressors.
In addition to the above synthetic nucleoside analogs, proteins, and antibodies, several plants and substances derived from plants have been found to have in vitro anti-HIV activity, such as Lonicera japonica and Prunella vulgaris, and glycyrrhizin from Glycyrrhiza radix. See R. S. Chang & H. W. Yeung, Antiviral Research, 9, 163-175 (1988) and M. Ito, et al., Antiviral Research, 7, 127-137 (1987).
Despite all of the available pharmaceuticals for the treatment of HIV, there is still no cure for the deadly disease. HIV viruses continue to mutate and become resistant to existing drugs such as the reverse transcriptase and protease inhibitors. Recently, a therapy of using two (2) or three (3) anti-HIV drugs in combination has been found effective in significantly lowering the HIV loads in AIDS patients. The results have been promising. However, the virus continues to develop resistance to the drugs and the long-term outcome (survival and cure rates) is still unknown. Thus, the medical communities throughout the world continue to search for drugs that can prevent the HIV infections, treat the HIV carriers to prevent them from progressing to full-blown deadly AIDS, and treat the AIDS patients.
Herbal Medicines
The use of herbal drugs and folk medicines have been known for thousands of years in China. These herbal approaches to the treatment of numerous illnesses, from arthritis to viral infections, have been previously viewed by western medicine as ineffective and dangerous. During the 19th century, many home remedies containing herbs were patented and sold. Modern drugs have replaced those remedies, but many modern drugs contain ingredients derived from herbs. For example, in 1776 the English botanist and physician William Withering learned that an herbal tea made by an old farm woman was effective in treating dropsy, or excess water in the tissues, which is caused by the inability of the heart to pump strongly enough. He found that one ingredient of the tea, which was made from leaves of the foxglove plant, strengthened the heart's pumping ability. The drug made from the foxglove plant is now known as digitalis.
Folk medicine is a relatively modern term to the West and has come to mean the care and treatment of the sick through a variety of herbal medicines. In recent years, folk medicines have become of increasing interest to many people in the western scientific medical community.
Prior Art--Herbal Medicines
A Chinese herbal medicine known as AEGINETIAE HERBA (a.k.a., GOLDEN LOCK KEY or LOTUS HERBA); has traditionally been used to treat illnesses such as swollen and sore throat, urinary tract infection, osteomyelitis, boil, tonsillitis, goiter, pharyngitis, thyroiditis, enteritis, liver disease, cancer, rheumatism, hematemesis, neurasthenia, eye redness, piles, menstruation irregularity, dropsy, jaundice, hernia, snake bite, and child developmental retardation. AEGINETIAE HERBA is prepared from the dried whole plant of Aeginelia indica which belongs to the family of Orobanchaceae. Dichondra micrantha, Striga lutea and Dichondra repens are also used to prepare this herbal medicine. Treatment dosage using the dried plant is typically from 4 to 150 g per day. It should be noted that the plant tastes bitter and is toxic.
Okubo et al. disclose that a phosphate buffered saline (PBS) extract (pH 7.2 at ambient to 4.degree. C.) from the seeds of Aeginetia indica exhibits excellent carcinostatic effect and possesses interleukin-2 and interferon-.gamma. inducing potency. The PBS was a 0.1 M phosphate buffered physiological saline at pH 7.2, not containing calcium or magnesium ions. The extracted substance is taught to be a macromolecular polysaccharide which may or may not contain lipid A binding with protein depending on whether the extraction is conducted using butanol or phenol. The extracted substance was soluble in water and insoluble in n-butanol. Its molecular weight was within the range of 100,000 to 200,000 dalton. See S. Okubo, M. Sato, & K. Himeno, U.S. Pat. No. 5,366,725, issued on Nov. 22, 1994.
A Chinese herbal medicine known as BAPHICACANTHIS RHIZOMA ET RADIX has traditionally been used to treat numerous illnesses such as fever, abscess, erysipelas, swollen sore throat, headache, jaundice, plague, leucorrhea, and syphilis. BAPHITCACANTHIS RHIZOMA ET RADIX is prepared from the dried rhizoma and root of Baphicacanthes cusia, Strobilanthes cusia, Isatis tinctoria, Isatis indigotica, or Polygonum tinctorium. It has been reported that this herbal medicine has exhibited inhibition of flu virus in vitro. A decoction from boiling the root of Isatis tinctoria in water has also exhibited antibacterial effect. Baphicacanthes cusia and Strobilanthes cusia belong to the family of Acanthaceae. Isatis tinctoria and Isatis indigotica belong to the family of Cruciferae. Polygonum tinctorium belongs to the family of Polygonaceae. Treatment doses are typically 10 to 19 g per day for BAPHICACANTHIS RHIZOMA ET RADIX.
Ho et al. disclose the use of an extract from a mixture of herbs including Isatis tinctoria for the in vitro inhibition of HIV infection in human T lymphocyte cells and mononuclear phagocytic lineage cells. The activity was based on the test results of a water extract from a mixture of three herbs: Isatis tinctoria (or Isatis indigotica), Lonicera japonica, and Polygonum bistorta. See D. D. Ho & X. S. Li, U.S. Pat. No. 5,178,865, issued on Jan. 12, 1993.
A compound known as tryptanthrin has been identified as the principal antifungal agent in the leaf of Strobilanthes cusia and as the main antidermatophytic substance in the leaf of Polygonum tinctorium and Isatis tinctoria. See H. Y. Hsu, Y. P. Chen, & M. Hong, The Chemical Constituents Of Oriental Herbs, Vol. 2, Oriental Healing Arts Institute, Los Angeles, Calif., U.S.A., 758-759 (1985).
The Chinese herbal medicine known as BLECHNI RHIZOMA, which is also known as DRYOPTERIS CRASSIRHIZOMAE RHIZOMA has traditionally been used to treat illnesses such as cut injury, swelling, fever, measles and erysipelas. BLECHNI RHIZOMA is prepared from the dried root and stem of Blechnum orientate which belongs to the family of Polypodiaceae or Blechnaceae. DRYOPTERIS CRASSIRHIZOMAE RHIZOMA is prepared from the dried root and stem of Dryopteris crassirhizoma which belongs to the family of Aspidiaceae. Osmunda japonica (Osmundaceae family), Woodwardia orientalis and Woodwardia unigemmata (Blechnaceae family), Athyrium acrostichoides (Aspidiaceae or Athyriaceae family), Sphaeropteris lepifera (Cyatheaceae family), Cyrtomium falcatum, and Cyrtomium fortunei (Aspidiaceae family) have also been used for preparation of these herbal medicines. These herbal medicines taste bitter and astringent and are slightly toxic. Treatment dosages are typically 4 to 11 g per day.
Blechnum orientate has also shown a strong inhibition effect against the influenza virus. Filmarone, filicin, aspidin, albaspidin, and filicic acid which are found in Dryopteris crassirhizoma have been characterized as having an anthelmintic effect. See H. Y. Hsu, Y. P. Chen, S. G. Hsu, J. S. Hsu, C. J. Chen, & H. C. Chang, Concise Pharmacognosy, New Medicine Publishing Co., Taipei, R.O.C., 577-578 (1985); and H. Y. Hsu, Y. P. Chen, & M. Hong, The Chemical Constituents Of Oriental Herbs, Oriental Healing Arts Institute, Los Angeles, Calif., U.S.A., 249-250 (1982).
Hozumi et al. disclose the rhizome of Dryopteris crassirhizoma as an antiherpesviral agent, antipolioviral agent, and anti-varicella-zoster virus agent. The rhizome of Cyrtomium fortunei and the rhizome of Woodwardia orientalis are also disclosed as antiherpesviral, antipolioviral, anti-measles virus, anti-varicella-zoster virus, and anti-cytomegalovirus (CMV) agents, as well as an anti-DNA and anti-RNA virus agent. See T. Hozumi, T. Matsumoto, H. Ooyama, T. Namba, K. Shiraki, M. Hattori, M. Kurokawa, & S. Kadota, U.S. Pat. No. 5,411,733, issued May 2, 1995.
A Chinese herbal medicine known as BLETILLAE TUBER has traditionally been used to treat illnesses such as hemoptysis, epistaxis, hematemesis, abscess, burn, dry and chapped skin, tuberculosis, gastric ulcer, and sores. BLETILLAE TUBER has astringent, antibacterial and antifungal properties. BLETILLAE TUBER is prepared from the dried tuber of Bletilla striata which belongs to the family of Orchidaceae. BLETILLAE TUBER tastes bittersweet, astringent and is nontoxic. Treatment dose is typically 2 to 11 g per day for an average human.
Chinese herbal medicines known as CIRSII RHIZOMA ET RADIX and BREEAE RADIX have traditionally been used to treat illnesses such as hematemesis, acute infectious hepatitis, cut bleeding, sores, and abscess. CIRSII RHIZOMA ET RADIX is prepared from the dried rhizoma or root or the whole plant of plants such as Cirsium japonicum, Cirsium albescens, and Cirsium japonicum var. australe which are from the Compositae family. BREEAE RADIX is prepared from the dried root of Compositae family plants such as Breea segetum (also known as Cephalanoplos segetum) and Breea setosum. Both herbal medicines taste sweet and slightly bitter, and are nontoxic. Treatment dose is typically 5 to 75 g per day for the average human.
A Chinese herbal medicine known as DICHONDRAE HERBA has traditionally been used to treat illnesses such as jaundice, dysentery, gonorrhea, dropsy, swollen boil, convulsion, encephalitis, rheumatism, hernia, diabetes mellitus, and hypertension. DICHONDRAE HERBA is prepared from the dried whole plant of Dichondra repens or Dichondra micrantha which belongs to the family of Convolvulaceae. The plant tastes bitter and is nontoxic. Treatment dosage of the dried plant is typically 10 to 40 g per day. Nine (9) compounds which were isolated from n-hexane and ethanol extracts of the whole herb of Dichondra micrantha have been identified. These compounds are maltol, umbelliferone, scopoletin, umbelliferone-7-O-glucopyranoside, scopolin, astragalin, isoquercitrin, kaempferol-3-O-rutinoside, and quercetin-3-O-rutinoside. See C.-J. Chou, L.-C. Lin, S.-Y. Hsu and C.-F. Chen, J. Chin. Med., 4(2), 143-149 (1993).
A Chinese herbal medicine known as FORSYTHIAE FRUCTUS has traditionally been used to treat illnesses such as sores, abscess, lymph node swelling, urethritis, and hypertension. It was also found to inhibit several bacteria and influenza virus. FORSYTHIAE FRUCTUS is prepared from the dried mature fruit of Forsythia suspensa, Forsythia viridissima, or Forsythia koreana which belong to the family Oleaceae. The herbal medicine tastes bitter and is nontoxic. Treatment dosage is typically 3 to 11 g per day.
Hozumi et al. disclose that the fruit of Forsythia suspensa is an antipolioviral agent and an anti-measles virus agent useful in treating these viral infections. See T. Hozumi, T. Matsumoto, H. Ooyama, T. Namba, K. Shiraki, M. Hattori, M. Kurokawa, & S. Kadota, U.S. Pat. No. 5,411,733, issued May 2, 1995.
The compounds forsythoside A (found in the leaf of Forsythia suspensa), forsythoside B (found in the stem of Forsythia koreana), and forsythoside C and forsythoside D (found in the fruit of Forsythia suspensa) have been reported to exhibit antibacterial activity against Staphylococcus aureus at a concentration less than 2 mM. Suspensaside (found in the fruit of Forsythia suspensa, likely the same as forsythoside C) has also been reported to exhibit antibacterial activity against Staphylococcus aureus Terashima with a minimum inhibition concentration (MIC) of 2.6 mg/mL. See H. Y. Hsu, T. P. Chen & M. Hong, The Chemical Constituents of Oriental Herbs, Vol. 2, Oriental Healing Arts Institute, Los Angeles, Calif., U.S.A., 53-55, 142-143 (1985).
A Chinese herbal medicine known as HEDYOTIS (also known as OLDENLANDIAE HERBA) has traditionally been used to treat illnesses such as urethra infection, pharyngitis, laryngitis, tonsillitis, subacute or chronic coccygodynia, appendicitis, intestinal cancer, contusion injury and eye disease. It has also been found to have weak antibacterial activity in vitro. HEDYOTIS is prepared from the dried whole plant of Hedyotis diffusa (also known as Oldenlandia diffusa) which belongs to the family Rubiaceae. The herbal medicine tastes sweet and is nontoxic. Treatment dosage is typically 19 to 300 g per day.
The Chinese herbal medicines known as LESPEDEZAE HERBA and SENECINIS HERBA have traditionally been used to treat illnesses such as urine incontinence, gonorrhea, asthma, stomach ache, general weakening and exhaustion, diarrhea, contusion injury, eye disease, eye redness, renal disease, acute inflammatory disease, cataract, dysentery, enteritis, jaundice, flu, septicemia, sore, swelling, and a disease of the palm. LESPEDEZAE HERBA is prepared from the dried whole plant of Lespedeza cuneata which belongs to the family Leguminosae. SENECINIS HERBA is prepared from the dried whole plant of Senecio scandens which belongs to the family Compositae. The extracts of Lespedeza cuneata and Senecio scandens have been shown to have an antibacterial effect. Both herbs taste sour, astringent and bitter. Treatment dose is typically 4 to 40 g per day.
A Chinese herbal medicine known as LIGUSTRI FRUCTUS has traditionally been used as a tonic and to treat illnesses such as insomnia, constipation, early white hair, neck lymph nodes tuberculosis and dropsy. LIGUSTRI FRUCTUS is prepared from the dried mature fruit of Ligustrum lucidum or Ligustrum japonicum which belongs to the family Oleaceae. The leaves of Ligustrum lucidum have been used as an antipyretic, analgesic, and anti-inflammatory agent. The leaves of Ligustrum japonicum have also been used to treat illnesses such as ophthalmalgia, ulcerative stomatitis, mastitis, swelling, and bum. The fruits of Ligustrum lucidum taste bitter and are nontoxic. Treatment dosage of the dried fruits is typically 6 to 20 g per day. That of the dried leaves is typically 40 to 75 g per day.
A Chinese herbal medicine known as LONICERAE FLOS has traditionally been used to treat illnesses such as fever, acute infectious diseases, measles, carbuncle, dysentery, enteritis, ringworm and similar skin diseases. LONICERAE FLOS is prepared from the dried flower bud of Lonicera japonica or Lonicera confusa. Both plants belong to the family Caprifoliaceae. The flower of Lonicera japonica has diuretic, antipyretic, anti-inflammatory, anti-convulsive, antibacterial and antiviral properties. The flower bud has also been used as a diuretic. The herbal medicine tastes sweet and is nontoxic. Treatment dosage is typically 11 to 75 g per day for the typical human.
Ho et al. disclose the anti-HIV activity in vitro of a mixture of Lonicera japonica, Isatis tinctoria (or Isatis indigotica) and Polygonum bistorta or a mixture of Lonicera japonica with Scutellaria baicalensis. Water extractions of the mixtures, treatment with ethanol precipitation and charcol adsorption are disclosed for the preparation of the anti-HIV active composition. See D. D. Ho & X. S. Li, U.S. Pat. No. 5,178,865, issued on Jan. 12, 1993. Several tannins such as caffeoylquinates isolated from Lonicera japonica have been reported to have an inhibitory effect on HIV-1 reverse transcriptase activity. See C.-W. Chang, M.-T. Lin, S.-S. Lee, K. C. S. C. Liu, F.-L. Hsu, & J.-Y. Lin, Antiviral Research, 27, 367-374 (1995).
A mixture of aqueous extracts of Lonicera japonica flower buds and Forsythia suspensa fruits with the crude flavonoids from Scutellaria baicalensis has been shown to have antibacterial and antiviral properties. A group of patients with severe respiratory disease were treated with the mixture and they responded as well as a control group on standard antibiotic therapy. See P. J. Houghton, Z. Boxu, & Z. Xisheng, Phytother. Res., 7, 384-386 (1993).
A Chinese herbal medicine known as PHELLODENDRI CORTEX has traditionally been used to treat illnesses such as dysentery, diarrhea, jaundice, stools with blood, abdominal pain, indigestion, bacteroid enteritis, and tuberculoid diarrhea. The herbal medicine has also been used to prepare an eye wash, for strengthening stomach and intestine to stimulate appetite, and as an astringent, anti-inflammatory, etc. It has antibacterial, anti-inflammatory, and wound healing properties. PHELLODENDRI CORTEX is prepared from the dried cortex of plants from the Rutaceae family such as Phellodendron amurense, Phellodendron chinense, Phellodendron amurense var. sachalinense, and Phellodendron wilsonii. PHELLODENDRI CORTEX tastes bitter and is nontoxic. Treatment dose is typically 1 to 11 g per day.
Hozumi et al. disclose the bark of Phellodendron amurense as antiherpesviral, antipolioviral, anti-measles virus, anti-varicella-zoster virus, and anti-CMV agents, as well as an anti-DNA virus and anti-RNA virus agent. See T. Hozumi, T. Matsumoto, H. Ooyama, T. Namba, K. Shiraki, M. Hattori, M. Kurokawa, & S. Kadota, U.S. Pat. No. 5,411,733, issued on May 2, 1995.
A Chinese herbal medicine known as POLYGONI CUSPIDATI RHIZOMA has traditionally been used to treat illnesses such as dysentery, menorrhagia, dysmenorrhea, dysuria, infantile growth retardation, and appendicitis. POLYGONI CUSPIDATI RHIZOMA is prepared from the dried rhizoma of Polygonum cuspidatum, Polygonum runcinatum, or Polygonum reynoutria (also known as Reynoutria japonica) which belong to the family Polygonaceae. The tender leaf has also been used to treat contusion and cut injuries. Extract of the herbal medicine has exhibited antibacterial and antiviral effects in vitro. Excessive use of the herbal medicine may cause a slight diarrhea. The herbal medicine tastes bitter and the treatment dose is typically 6 to 40 g per day.
Hozumi et al. disclose the root and rhizome of Polygonum cuspidatum as an antiherpesviral, antipolioviral, anti-varicella-zoster virus, and anti-CMV agent. See T. Hozumi, T. Matsumoto, H. Ooyama, T. Namba, K. Shiraki, M. Hattori, M. Kurokawa, & S. Kadota, U.S. Pat. No. 5,411,733, issued on May 2, 1995.
Resveratrol has been reported as an antifungal and antibacterial component in the root of Polygonum cuspidatum. See H. Y. Hsu, Y. P. Chen, & M. Hong, The Chemical Constituents Of Oriental Herbs, Vol. 2, Oriental Healing Arts Institute, Los Angeles, Calif., U.S.A., 51 (1985).
A Chinese herbal medicine known as PRUNELLAE SPICA has traditionally been used to treat illnesses such as goiter, hemorrhoids, swollen eye, ophthalmalgia, gonorrhea, uterine disease, mastitis, breast abscess, breast cancer, chronic arthritis, conjunctivitis, and hypertension. PRUNELLAE SPICA is prepared from the dried spica or whole plant of Prunella vulgaris or Prunella vulgaris subsp. asiatica (also known as Prunella vulgaris var. lilachina). Both plants belong to the family Labiatae. The whole plant can be used as a diuretic and also has antibacterial effect in vitro. The herbal medicine tastes bitter and is nontoxic. Treatment dosage is typically 4 to 110 g per day for the average human.
Hozumi et al. disclose the spike of Prunella vulgaris as an antiherpesviral agent for treating herpes virus infection. See T. Hozumi, T. Matsumoto, H. Ooyama, T. Namba, K. Shiraki, M. Hattori, M. Kurokawa, & S. Kadota, U.S. Pat. No. 5,411,733, issued May 2, 1995. The water extract of Prunella vulgaris (boiling 3 g in 100 mL water for 45 minutes) has also been reported to have anti-HIV (strain H9/3B) activity. The extract also exhibited synergistic anti-HIV activity with zidovudine (AZT) and didanosine (ddI). Only a slight additive effect was observed for Prunella vulgaris and zalcitabine (ddC). See J. F. John, R. Kuk, & A. Rosenthal, Abstr. Gen. Meet. Am. Soc. Microbiol., 94, 481(1994).
Yamasaki et al. evaluate in vitro two hundred four (204) crude drugs of common use in Japan for anti-HIV-1 activity and reported that the hot water extract of Prunella vulgaris (spike) showed a strong in vitro anti-HIV-1 activity with an IC.sub.100 of 16 .mu.g/mL. See K. Yamasaki, T. Otake, H. Mori, M. Morimoto, N. Ueba, Y. Kurokawa, K. Shiota, & T. Yuge, Yakugaku Zasshi, 113(11), 818-824 (1993).
Yao et al. reported that the water extract of the dried entire plant of Prunella vulgaris was active in vitro in inhibiting HIV-1 replication with relatively low cytotoxicity towards the MT-4 cells. The extract was also active in reverse transcriptase inhibition. The active factor was purified and identified as anionic with a molecular weight of approximately 10,000 dalton. This active component may be the same as the prunellin, as described below by Tabba, et al. The purified extract inhibited HIV-1 replication in the lymphoid cell line MT-4, in the monocytoid cell line U937, and in peripheral blood mononuclear cells (PBMC) at effective concentrations of 6, 30, and 12.5 .mu.g/mL, respectively. Pretreatment of uninfected cells with the extract prior to viral exposure did not prevent HIV-1 infection. Preincubation of HIV-1 with the purified extract dramatically decreased infectiousness. The purified extract was also able to block cell-to-cell transmission of HIV-1, prevented syncytium formation, and interfered with the ability of both HIV-1 and purified gp120 to bind to CD4. PCR (polymerase chain reaction) analysis confirmed the absence of HIV-1 proviral DNA in cells exposed to virus in the presence of the extract. The results suggested that the purified extract antagonized HIV-1 infection of susceptible cells by preventing viral attachment to the CD4 receptor. See X. J. Yao, M. A. Wainberg, & M. A. Parniak, Virology, 187(1), 56-62 (1992).
Tabba et al. isolated and partially characterized an anti-HIV component, prunellin, from aqueous extracts of dried inflorescence of Prunella vulgaris. Prunellin is a carbohydrate with an MIC (minimum inhibition concentration) of 2.2 .mu.g/mL against HIV-1 in vitro. It was identified as a partially sulfated polysaccharide with a molecular weight of about 10,000 dalton. See H. D. Tabba, R. S. Chang, & K. M. Smith, Antiviral Research, 11, 263-273 (1989).
Zheng evaluated four hundred seventy two (472) traditional medicinal herbs for antiviral effect on type 1 herpes simplex virus (HSV 1). Prunella vulgaris was one of the ten herbs found to be highly effective in vitro. Clinically, 78 cases of herpetic keratitis due to HSV1 were treated with Prunella vulgaris and Pyrrosia lingua eye drops. Among them, 38 cases were effectively cured, 37 cases showed an improvement, and 3 cases showed no benefit. See M. Zheng, J. Tradit. Chin. Med., 8(3), 203-206 (1988).
Triterpene 1 and triterpene 2 which have been isolated from Prunella vulgaris have shown antiviral activity against HSV1. Triterpene 1 was identified as betulinic acid and triterpene 2 was identified as 2.alpha., 3.alpha.-dihydroxyurs-12-en-28-oic acid. The EC.sub.50 was estimated to be 30 .mu.g/mL for triterpene 1 and 8 .mu.g/mL for triterpene 2 by plaque reduction assay. See S. Y. Ryu, C-K. Lee, C. O. Lee, H. S. Kim, & O. P. Zee, Arch. Pharmacal Res. (Seoul), 15(3), 242-245 (1992).
A Chinese herbal medicine known as SCUTELLARIAE BARBATAE HERBA has traditionally been used to treat illnesses such as hematemesis, gonorrhea with traces of blood, sores, cancer, convulsion, pneumonia, enteritis, coccygodynia, appendicitis, asthma, malaria, and rheumatism. It was also found to have antibacterial effect. SCUTELLARIAE BARBATAE HERBA is prepared from the dried whole plant of Scutellaria barbata, Scutellaria rivularis, or Scutellaria dependens which belong to the family Labiatae. The herbal medicine tastes bitter and should not be consumed by those who have anemia. Pregnant women should avoid taking this herb. Treatment dosage is typically 4 to 300 g per day.
Dried whole plants of Scutellaria rivularis have been used in folk medicine for the treatment of tumors, hepatitis, liver cirrhosis, and other diseases in China and Taiwan. See Y. L. Lin, Y. H. Kuo, G. H. Lee, and S. M. Peng, J. Chem. Research (S), 320-321 (1987).
Apigenin, isolated from the whole herb of Scutellaria rivularis, was found to have anti-influenza virus activity. See T. Nagai, et al., Chem. Pharm. Bull., 38(5), 1329-1332 (1990).
A Chinese herbal medicine known as SOLANI HERBA has traditionally been used to treat illnesses such as boil, acute nephritis, cancer and sores. SOLANI HERBA is prepared from the dried whole plant of Solanum nigrum which belongs to the family Solanaceae. The extract of SOLANI HERBA has demonstrated anti-inflammatory property. The fruit has also exhibited the effects of suppressing coughs and relieving bronchial inflammation. The herbal medicine tastes bitter and slightly sweet, and is nontoxic. Treatment dosage is typically 11 to 60 g per day.
The compound solasonine (found in the whole herb, fruit, leaf, and fresh immature berries of Solanum nigrum) has an anti-inflammatory effect similar to cortisone. Solasonine and solanine (also found in Solanum nigrum) possess the ability of raising or lowering the blood sugar level in rats depending on the situation of the animals. Solasonine was also reported to have a stimulating effect on the heart, while solanine had a suppressive effect. When administered at small doses, solasonine enhances the stimulative process of the central nerve system in animals (i.e., rat and rabbit). On the other hand, it enhances the suppressive process when administered at large doses. Solasonine can also lower the blood coagulability. See (1) H. Y. Hsu, Y. P. Chen, S. G. Hsu, J. S. Hsu, C. J. Chen, & H. C. Chang, Concise Pharmacognosy, New Medicine Publishing Co., Taipei, R.O.C., 176-177 (1985); (2) H. Y. Hsu, Y. P. Chen, & M. Hong, The Chemical Constituents Of Oriental Herbs, Oriental Healing Arts Institute, Los Angeles, Calif., U.S.A., 1400-1401, 1406 (1982); and (3) H. Y. Hsu, Y. P. Chen, & M. Hong, The Chemical Constituents Of Oriental Herbs, Vol. 2, Oriental Healing Arts Institute, Los Angeles, Calif., U.S.A., 742 (1985).
Combinations of herbal medicines such as LONICERAE FLOS, BAPHICACANTHIS RHIZOMA ET RADIX, and FORSYTHIAE FRUCTUS have been used as antipyretic and detoxification agents and for treating acute hepatitis. The herbal medicines BLECHNI RHIZOMA and POLYGONI CUSPIDATI RHIZOMA have been used along with other herbal medicines in a formula for treating Hepatitis B. The herbal medicines SCUTELLARIAE BARBATAE HERBA and LIGUSTRI FRUCTUS have occasionally been added with other herbal medicines into the above formula to improve the treatment. The herbal medicine LIGUSTRI FRUCTUS was occasionally used along with other herbal medicines mainly as a tonic and the herbal medicine HEDYOTIS has been occasionally used along with other herbal medicines as a detoxification agent. The herbal medicine PRUNELLAE SPICA has also been used along with other herbal medicines to relief liver stress.
Chang and Yeung screened the boiling water extracts of twenty seven (27) medicinal herbs for anti-HIV activity. They found eleven (11) of the extracts were active in inhibiting HIV in the H9 cells. Lonicera japonica, Prunella vulgaris, Woodwardia unigemmata, and Senecio scandens were among those active ones with moderate activities. Forsythia suspensa, Isatis tinctoria, and Polygonum cuspidatum were among those tested which did not display activity in the anti-HIV assay. The anti-HIV active extract of Viola yedoensis was further tested and found to be fairly specific. The extract did not inactivate HIV extracellularly and did not inhibit the growth of herpes simplex, polio, or vesicular stomatitis viruses in human fibroblast culture. See R. S. Chang & H. W. Yeung, Antiviral Research, 9, 163-175 (1988).
Antiviral agents have been isolated from Syzygium aromaticum, Sapium sebiferum, Scutellaria baicalensis, and Scutellaria rivularis. Eugeniin (a tannin) isolated from Syzygium aromaticum and methyl gallate isolated from Sapium sebiferum exhibited anti-herpes simplex virus activity in vitro. Plant flavenoids, such as 5,7,4'-trihydroxy-8-methoxyflavone from the root of Scutellaria baicalensis and apigenin (5,7,4'-trihydroxyflavone) from the whole herb Scutellaria rivularis, were also reported to have anti-influenza virus activity. See (1) T. Hozumi, et al., U.S. Pat. No. 5,411,733 (1995); (2) M. Takechi & Y. Tanaka, Planta Medica, 42, 69-74 (1981); (3) C. J. M. Kane, et al, Bioscience Reports, 8, 85-94 (1988); and (4) T. Nagai, et al., Chem. Pharm. Bull., 38(5), 1329-1332 (1990).
Hozumi et al. investigated ninety one (91) herbal medicines which demonstrated antiviral activity. More specifically, fifty two (52) of them had antiherpesviral activity, sixty four (64) had antipolioviral activity, thirty seven (37) had anti-measles virus activity, twenty seven (27) had anti-varicella-zoster virus activity, twenty three (23) had anti-CMV activity, and twenty eight (28) had anti-DNA virus and anti-RNA virus activity. See T. Hozumi, T.
Matsumoto, H. Ooyama, T. Namba, K. Shiraki, M. Hattori, M. Kurokawa, & S. Kadota, U.S. Pat. No. 5,411,733, issued on May 2, 1995. The anti-DNA virus and anti-RNA virus activity of the twenty eight (28) herbal medicines disclosed in the 5,411,733 patent was based upon their antiherpesviral, antipolioviral, anti-measles virus, and/or anti-varicella-zoster virus and anti-CMV activities. However, the extrapolation to cover both anti-DNA virus and anti-RNA virus activities is unfounded from the experiments conducted.
The data of the present invention, presented below, evidenced little or no anti-HIV activity of the two herbal medicines at 2.5 and 0.5 mg/mL derived from the rhizome of Cyrtomium fortunei and the bark of Phellodendron amurense. In contrast, the three (3) herbal medicines using the spike of Prunella vulgaris, the fruit of Forsythia suspensa, and the root and rhizome of Polygonum cuspidatum will be shown to have a strong to moderate anti-HIV activity at 2.5 mg/mL. Prunella vulgaris has also been reported by others as described above to have a very good anti-HIV activity.
It is noted that in the practice of traditional Chinese medicine, herbal medicines are used to treat the symptoms of the patients, not the disease or causative agent itself, and are therefore not known to be specific to a particular disease. Herbal medicines were prescribed depending on the symptoms of the individual patient. The composition of herbal medicines also vary case by case and may even change for each individual patient during the course of the treatment according to each treatment result. It is therefore very difficult to describe a particular herbal composition from the prior art suitable for treating a specific disease.
The present invention is directed to the discovery of antiviral herb compositions, extracts thereof and the active chemical constituents thereof. The antiviral herb compositions of this invention are derived from individual herbs, herb mixtures and commercially available Chinese herbal medicines. These novel herb compositions and their extracts and/or active principles are demonstrated herein as active against viral diseases such as HBV and HCV carriers, hepatitis B, hepatitis C, HIV infection and AIDS.